Back to Search
Start Over
USP28 and USP25 are downregulated by Vismodegib in vitro and in colorectal cancer cell lines.
- Source :
-
The FEBS journal [FEBS J] 2021 Feb; Vol. 288 (4), pp. 1325-1342. Date of Electronic Publication: 2020 Jul 20. - Publication Year :
- 2021
-
Abstract
- Deubiquitinase USP28 plays a crucial role in tumorigenesis by enhancing the stabilities of multiple cancer-related proteins including c-Myc, Notch1, and LSD1, and has become an attractive target for anticancer drug development. However, to date, only a few of USP28-targeted active compounds have been developed, and the active compound-binding pocket in USP28 has not been experimentally revealed yet. In this study, bioassay-based high-throughput screening was applied to discover USP28-targeted inhibitors from the commercially available drug library. Vismodegib, an inhibitor of Hedgehog signaling pathway and FDA-approved drug for the treatment of basal cell carcinoma, was found to exhibit inhibition activity against USP28 (IC <subscript>50</subscript> : 4.41 ± 1.08 μm). Multiple biophysical and biochemical techniques including NMR, ITC, thermal shift assay, HDX-MS, and site-directed mutagenesis analysis were then used to characterize the interaction between Vismodegib and USP28. The binding pocket in USP28 for Vismodegib, which is mainly composed of two helical structures spanning D255-N278 and N286-Y293, was revealed. According to the possible binding pose generated by HDX-MS data-defined molecular docking, the binding cavity occupied by Vismodegib in USP28 aligns well with one of the reported-binding pockets in USP7 for its inhibitors. Furthermore, cellular assays were conducted to confirm that Vismodegib could interact with the evolutionarily related deubiquitinases USP28 and USP25 and downregulate the levels of the two enzymes' substrate proteins c-Myc, Notch1, and Tankyrase-1/2.<br /> (© 2020 Federation of European Biochemical Societies.)
- Subjects :
- Anilides chemistry
Anilides metabolism
Biocatalysis drug effects
Carcinoma, Basal Cell drug therapy
Carcinoma, Basal Cell pathology
Cell Line, Tumor
Cell Survival drug effects
Colorectal Neoplasms drug therapy
Colorectal Neoplasms pathology
HCT116 Cells
Hedgehog Proteins metabolism
Humans
Molecular Docking Simulation
Molecular Structure
Protein Binding
Protein Domains
Pyridines chemistry
Pyridines metabolism
Signal Transduction drug effects
Ubiquitin Thiolesterase antagonists & inhibitors
Ubiquitin Thiolesterase chemistry
Anilides pharmacology
Carcinoma, Basal Cell metabolism
Colorectal Neoplasms metabolism
Down-Regulation drug effects
Pyridines pharmacology
Ubiquitin Thiolesterase metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1742-4658
- Volume :
- 288
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- The FEBS journal
- Publication Type :
- Academic Journal
- Accession number :
- 32578360
- Full Text :
- https://doi.org/10.1111/febs.15461